Activation and Identification of a Griseusin Cluster in <i>Streptomyces</i> sp. CA-256286 by Employing Transcriptional Regulators and Multi-Omics Methods

<i>Streptomyces</i> are well-known producers of a range of different secondary metabolites, including antibiotics and other bioactive compounds. Recently, it has been demonstrated that “silent” biosynthetic gene clusters (BGCs) can be activated by heterologously expressing transcriptiona...

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Autores principales:	Charlotte Beck, Tetiana Gren, Francisco Javier Ortiz-López, Tue Sparholt Jørgensen, Daniel Carretero-Molina, Jesús Martín Serrano, José R. Tormo, Daniel Oves-Costales, Eftychia E. Kontou, Omkar S. Mohite, Erik Mingyar, Evi Stegmann, Olga Genilloud, Tilmann Weber
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	transcriptional regulators biosynthetic gene cluster genome mining heterologous expression griseusin pyranonaphtoquinone Organic chemistry QD241-441
Acceso en línea:	https://doaj.org/article/86badcd3f9eb4ba6bb5883784ca814da
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spelling	oai:doaj.org-article:86badcd3f9eb4ba6bb5883784ca814da2021-11-11T18:33:36ZActivation and Identification of a Griseusin Cluster in <i>Streptomyces</i> sp. CA-256286 by Employing Transcriptional Regulators and Multi-Omics Methods10.3390/molecules262165801420-3049https://doaj.org/article/86badcd3f9eb4ba6bb5883784ca814da2021-10-01T00:00:00Zhttps://www.mdpi.com/1420-3049/26/21/6580https://doaj.org/toc/1420-3049<i>Streptomyces</i> are well-known producers of a range of different secondary metabolites, including antibiotics and other bioactive compounds. Recently, it has been demonstrated that “silent” biosynthetic gene clusters (BGCs) can be activated by heterologously expressing transcriptional regulators from other BGCs. Here, we have activated a silent BGC in <i>Streptomyces</i> sp. CA-256286 by overexpression of a set of SARP family transcriptional regulators. The structure of the produced compound was elucidated by NMR and found to be an <i>N</i>-acetyl cysteine adduct of the pyranonaphtoquinone polyketide 3′-<i>O</i>-α-<span style="font-variant: small-caps;">d</span>-forosaminyl-(+)-griseusin A. Employing a combination of multi-omics and metabolic engineering techniques, we identified the responsible BGC. These methods include genome mining, proteomics and transcriptomics analyses, in combination with CRISPR induced gene inactivations and expression of the BGC in a heterologous host strain. This work demonstrates an easy-to-implement workflow of how silent BGCs can be activated, followed by the identification and characterization of the produced compound, the responsible BGC, and hints of its biosynthetic pathway.Charlotte BeckTetiana GrenFrancisco Javier Ortiz-LópezTue Sparholt JørgensenDaniel Carretero-MolinaJesús Martín SerranoJosé R. TormoDaniel Oves-CostalesEftychia E. KontouOmkar S. MohiteErik MingyarEvi StegmannOlga GenilloudTilmann WeberMDPI AGarticletranscriptional regulatorsbiosynthetic gene clustergenome miningheterologous expressiongriseusinpyranonaphtoquinoneOrganic chemistryQD241-441ENMolecules, Vol 26, Iss 6580, p 6580 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	transcriptional regulators biosynthetic gene cluster genome mining heterologous expression griseusin pyranonaphtoquinone Organic chemistry QD241-441
spellingShingle	transcriptional regulators biosynthetic gene cluster genome mining heterologous expression griseusin pyranonaphtoquinone Organic chemistry QD241-441 Charlotte Beck Tetiana Gren Francisco Javier Ortiz-López Tue Sparholt Jørgensen Daniel Carretero-Molina Jesús Martín Serrano José R. Tormo Daniel Oves-Costales Eftychia E. Kontou Omkar S. Mohite Erik Mingyar Evi Stegmann Olga Genilloud Tilmann Weber Activation and Identification of a Griseusin Cluster in <i>Streptomyces</i> sp. CA-256286 by Employing Transcriptional Regulators and Multi-Omics Methods
description	<i>Streptomyces</i> are well-known producers of a range of different secondary metabolites, including antibiotics and other bioactive compounds. Recently, it has been demonstrated that “silent” biosynthetic gene clusters (BGCs) can be activated by heterologously expressing transcriptional regulators from other BGCs. Here, we have activated a silent BGC in <i>Streptomyces</i> sp. CA-256286 by overexpression of a set of SARP family transcriptional regulators. The structure of the produced compound was elucidated by NMR and found to be an <i>N</i>-acetyl cysteine adduct of the pyranonaphtoquinone polyketide 3′-<i>O</i>-α-<span style="font-variant: small-caps;">d</span>-forosaminyl-(+)-griseusin A. Employing a combination of multi-omics and metabolic engineering techniques, we identified the responsible BGC. These methods include genome mining, proteomics and transcriptomics analyses, in combination with CRISPR induced gene inactivations and expression of the BGC in a heterologous host strain. This work demonstrates an easy-to-implement workflow of how silent BGCs can be activated, followed by the identification and characterization of the produced compound, the responsible BGC, and hints of its biosynthetic pathway.
format	article
author	Charlotte Beck Tetiana Gren Francisco Javier Ortiz-López Tue Sparholt Jørgensen Daniel Carretero-Molina Jesús Martín Serrano José R. Tormo Daniel Oves-Costales Eftychia E. Kontou Omkar S. Mohite Erik Mingyar Evi Stegmann Olga Genilloud Tilmann Weber
author_facet	Charlotte Beck Tetiana Gren Francisco Javier Ortiz-López Tue Sparholt Jørgensen Daniel Carretero-Molina Jesús Martín Serrano José R. Tormo Daniel Oves-Costales Eftychia E. Kontou Omkar S. Mohite Erik Mingyar Evi Stegmann Olga Genilloud Tilmann Weber
author_sort	Charlotte Beck
title	Activation and Identification of a Griseusin Cluster in <i>Streptomyces</i> sp. CA-256286 by Employing Transcriptional Regulators and Multi-Omics Methods
title_short	Activation and Identification of a Griseusin Cluster in <i>Streptomyces</i> sp. CA-256286 by Employing Transcriptional Regulators and Multi-Omics Methods
title_full	Activation and Identification of a Griseusin Cluster in <i>Streptomyces</i> sp. CA-256286 by Employing Transcriptional Regulators and Multi-Omics Methods
title_fullStr	Activation and Identification of a Griseusin Cluster in <i>Streptomyces</i> sp. CA-256286 by Employing Transcriptional Regulators and Multi-Omics Methods
title_full_unstemmed	Activation and Identification of a Griseusin Cluster in <i>Streptomyces</i> sp. CA-256286 by Employing Transcriptional Regulators and Multi-Omics Methods
title_sort	activation and identification of a griseusin cluster in <i>streptomyces</i> sp. ca-256286 by employing transcriptional regulators and multi-omics methods
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/86badcd3f9eb4ba6bb5883784ca814da
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Activation and Identification of a Griseusin Cluster in <i>Streptomyces</i> sp. CA-256286 by Employing Transcriptional Regulators and Multi-Omics Methods

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